Type II collagen quantification in experimental chondrogenesis

O’Driscoll, Shawn W.; Commisso, Cinzia N.; Fitzsimmons, James S.

doi:10.1016/s1063-4584(05)80054-8

Cited by 34 publications

(25 citation statements)

References 26 publications

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“…A number of methods have been developed to determine the proportion of collagen types in various tissues, particularly type-I, type-II and type-III collagens. 28,31,32 These generally involve isolation of the collagen fraction of the extracellular matrix, followed by chromatographic separation of cyanogen-bromide-digested peptides (CB-peptides). They are highly specific and each collagen type isolated to date has its own unique CB-peptide pattern.…”

Section: Discussionmentioning

confidence: 99%

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Chondrocyte Transplantation Using a Collagen Bilayer Matrix for Cartilage Repair

Frenkel

Toolan

Menche

et al. 1997

The Journal of Bone and Joint Surgery. British volume

105

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We have developed a novel, two-layered, collagen matrix seeded with chondrocytes for repair of articular cartilage. It consists of a dense collagen layer which is in contact with bone and a porous matrix to support the seeded chondrocytes. The matrices were implanted in rabbit femoral trochleas for up to 24 weeks. The control groups received either a matrix without cells or no implant.The best histological repair was seen with cell-seeded implants. The permeability and glycosaminoglycan content of both implant groups were nearly normal, but were significantly less in tissue from empty defects. The type-II collagen content of the seeded implants was normal. For unseeded implants it was 74.3% of the normal and for empty defects only 20%. The current treatments for articular injury often result in a fibrous repair which deteriorates with time. This bilayer implant allowed sustained hyaline-like repair of articular defects during the entire six-month period of observation. [Br] 1997;79-B:831-6. Received 23 September 1996; Accepted after revision 18 April 1997 The clinical need for repair of lesions of the articular cartilage is increasing, but there is no satisfactory method of surgical repair. Abrasion arthroplasty, excision and drilling, cartilage debridement, and arthroscopic shaving have all been used. J Bone Joint Surg

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Section: Discussionmentioning

confidence: 99%

“…Histological specimens were scored blindly by a grading system modified from O'Driscoll, Comisso and Fitzsimmons. 28 The criteria included the percentage of hyaline cartilage, surface regularity, thickness, cellularity, cluster formation, bonding to native cartilage, and degeneration of adjacent cartilage. The maximum score was 24 points.…”

Section: Methodsmentioning

confidence: 99%

Chondrocyte Transplantation Using a Collagen Bilayer Matrix for Cartilage Repair

Frenkel

Toolan

Menche

et al. 1997

The Journal of Bone and Joint Surgery. British volume

105

View full text Add to dashboard Cite

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“…To confirm the histology results, quantitative collagen typing was performed [33]. After six weeks in culture, quantitative collagen typing was performed using a published technique for measuring the relative amount of type I1 collagen with respect to type I collagen in tissue samples [44].…”

Section: Collugen Typingmentioning

confidence: 99%

“…After six weeks in culture, quantitative collagen typing was performed using a published technique for measuring the relative amount of type I1 collagen with respect to type I collagen in tissue samples [44]. This technique has been modified to permit the analysis of very small samples (1-10 mg) without initial purification of the collagen [33]. Samples were weighed, and the collagen peptides were cleaved with 0.5 ml 5% cyanogen bromide (CNBr) in deaerated 88% formic acid.…”

Section: Collugen Typingmentioning

confidence: 99%

Serum‐free media for periosteal chondrogenesis in vitro

Fitzsimmons

Sanyal

Gonzalez

et al. 2004

Journal Orthopaedic Research

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Organ culture studies involving whole explants of periosteum have been useful for studying chondrogenesis, but to date the standard culture model for these explants has required the addition of fetal bovine serum to the media. Numerous investigators have succeeded in culturing chondrocytes and embryonic cells in serum-free conditions but there have been no studies focused on achieving a defined, serum-free media for culturing periosteal explants. The purpose of the present investigation was to determine if whole periosteal explants can be grown and produce cartilage in serum-free conditions, and to define the minimum media supplements that would be conducive to chondrogenesis. 321 periosteal explants were obtained from the medial proximal tibiae of 31 two month-old NZ white rabbits and cultured using a published agarose suspension organ culture model and DMEM for six weeks. The explants were cultured with and without fetal bovine serum or bovine serum albumin and exposed to transforming growth factor beta alone, a combination of growth factors we call ChondroMix (10 ng/ml transforming growth factor beta, 50 ng/ml basic fibroblast growth factor, and 5 pg/ml growth hormone), and/or ITS+ (2.08 pg/ml each of insulin, transferrin, and selenious acid, plus 1.78 pg/ml linoleic acid and 0.42 mg/ml BSA). Maximal chondrogenic stimulation in this study was observed with the combination of ChondroMix and ITS+. However, the minimal requirement to match or exceed the level of chondrogenic stimulation seen in the standard model (TGF-1 in 100/0 FBS) was achieved simply by the addition of 2.0 pg/ml insulin in 0.10/0 BSA-containing medium (p < 0.05). Therefore, based on our results, it would be reasonable to assume that insulin is the component in ITS+ responsible for the observed increase in total cartilage growth. Lower concentrations of insulin were not effective, suggesting that the observed effect of insulin requires activation of the IGF-1 receptor.

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“…This technique has been modilied to permit the analysis of bery small samples ( 1 -10 pg) without initial purification of the collagen [24]. Samples were weighed for wet weight.…”

Section: Sl~i~/l~llll Fll'/rllrir~mentioning

confidence: 99%

The chondrogenic potential of periosteum decreases with age

O’Driscoll

Saris

Ito

et al. 2001

Journal Orthopaedic Research

166

121

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Periosteum contains undifferentiated inesenchymal stem cells that possess the potential for chondrogenesis during cartilage repair and in fracture healing. With aging, the chondrogenic potential of periosteum declines significantly. An organ-culture model was used to investigate the relationship between the chondrogenic potential of periosteum and aging. A total of 736 periosteal explants from the proximal medial tibiae of 82 rabbits, aged 2 weeks to 2 years, were cultured in agarose suspension conditions conductive for chondrogenesis, and analyzed using histomorphometry, collagen typing, wet weight measurement, jH-thymidine and "S-sulfate uptake, autoradiography, and PCNA immunostaining. The rabbits were skeletally mature by 6 months and stopped increasing in weight by 12 months. Chondrogenesis declined significantly with age ( P < 0.0001) and was maximal in the 1.5-2 month-old rabbits, Explants from the 6 month-old rabbits formed 50% less cartilage, and by 12 months chondrogenesis reached a steady state minimal level. In parallel with this decrease in chondrogenic potential similar decreases were measured in 'H-thymidine uptake ( P < 0.0001 ). "S-sulfate uptake ( P = 0.01 17), as well as the thickness ( P < 0.0001) and the total number of cells in the cambium layer of the periosteum ( P < 0.0001 ). Autoradiography with 'H-thymidine and PCNA immunostaining confirmed the measured decrease in proliferative activity in the cambium layer where the chondrocyte precursors reside, although the percentage of proliferating cells did not change significantly with age. The most dramatic change was the marked decrease (87"h) in the thickness and total cell number in the cambium layer of the perisoteum between the 2 and 12 month-old rabbits ( P < 0.05). These data confirm a decline in the chondrogenic potential of periosteum with aging. Thus, one possibility for improving cartilage formation by periosteal transplantation after skeletal maturity would be to stimulate an increase in the total number of cells in the chondrocyte precursor pool early during chondrogenesis.

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Type II collagen quantification in experimental chondrogenesis

Cited by 34 publications

References 26 publications

Chondrocyte Transplantation Using a Collagen Bilayer Matrix for Cartilage Repair

Chondrocyte Transplantation Using a Collagen Bilayer Matrix for Cartilage Repair

Serum‐free media for periosteal chondrogenesis in vitro

The chondrogenic potential of periosteum decreases with age

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